Skip to main content

Advertisement

SpringerLink
Log in

Regulatory T cells are associated with post-cryoablation prognosis in patients with hepatitis B virus-related hepatocellular carcinoma

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Background

We carried out this study to evaluate the association between regulatory T cells (Treg) and prognosis and progression after cryoablation in patients with hepatitis-B virus-related hepatocellular carcinoma.

Methods

Peripheral Treg frequency in 111 patients with hepatocellular carcinoma (HCC) was detected by flow cytometry. Treg frequency and function were re-examined during patient follow up. A possible association between Treg and α-fetoprotein (AFP) was also analyzed, and the distribution of resident CD4+ and CD8+ T cells and FoxP3+ T cells in the liver tissue of patients with HCC was examined by immunohistochemistry.

Results

Treg frequency significantly increased with disease progression. Our longitudinal study showed that Treg frequency had significantly decreased in 17 patients with HCC regression following cryoablation, but the frequency had dramatically increased in 14 patients with HCC recurrence or progression. Furthermore, AFP levels varied in a way comparable with Treg frequency in patients with elevated AFP recorded before therapy. Significantly increased suppressive effects of Treg on proliferation and cytokine secretion of CD8+ and CD4+ T cells were observed during follow up in patients with tumor progression, but not in patients with tumor response. Moreover, the numbers of CD8+, CD4+, and FoxP3+ cells infiltrating the tumors around the cryotherapeutic zones were significantly decreased after argon–helium cryoablation, and this was associated with a reduction in the FoxP3/CD8 ratio. Importantly,increased quantities of circulating CD4+CD25+FoxP3+ Treg and tumor infiltrating FoxP3+ cells before cryoablation were associated with high recurrence or risk of progression in HCC patients after cryoablation.

Conclusions

Treg variation is associated with tumor regression or progression in HCC following cryoablation and may be used as a marker to estimate HCC progression.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

APC :

Allophycocyanin

PerCP:

Peridinin chlorophyll protein

FITC :

Fluorescein isothiocyanate

PE :

Phycoerythrin

Treg :

Regulatory T cells

AFP :

α-Fetoprotein

PBMC :

Peripheral blood mononuclear cell

HCC :

Hepatocellular carcinoma

TR :

Tumor response

TP :

Tumor progression

References

  1. Parkin DM, Bray F, Ferlay J, Pisani P. Estimating the world cancer burden: Globocan 2000. Int J Cancer. 2001;94:153–6.

    Article  CAS  PubMed  Google Scholar 

  2. Jemal A, Murray T, Ward E, Samuels A, Tiwari RC, Ghafoor A, et al. Cancer Statistics, 2005. CA Cancer J Clin. 2005;55:10–30.

    Article  PubMed  Google Scholar 

  3. Gage AA. History of cryosurgery. Semin Surg Oncol. 1998;14:99–109.

    Article  CAS  PubMed  Google Scholar 

  4. Zhou L, Yang YP, Feng YY, Lu YY, Wang CP, Wang XZ, et al. Efficacy of argon-helium cryosurgical ablation on primary hepatocellular carcinoma: a pilot clinical study. AiZheng. 2009;28:45–8.

    Google Scholar 

  5. Yantorno C, Soanes WA, Gonder MJ, Shulman S. Studies in cryo-immunology I: the production of antibodies to urogenital tissue in consequence of freezing treatment. Immunology. 1967;12:395–410.

    CAS  PubMed  Google Scholar 

  6. Shulman S, Brandt EJ, Yantorno C. Studies in cryo-immunology. II: tissue and species specificity of the autoantibody response and comparison with iso-immunization. Immunology. 1968;14:149–58.

    CAS  PubMed  Google Scholar 

  7. Sabel MS, Nehs MA, Su G, Lowler KP, Ferrara JL, Chang AE. Immunologic response to cryoablation of breast cancer. Breast Cancer Res Treat. 2005;90:97–104.

    Article  CAS  PubMed  Google Scholar 

  8. Sabel MS, Arora A, Su G, Chang AE. Adoptive immunotherapy of breast cancer with lymph node cells primed by cryoablation of the primary tumor. Cryobiology. 2006;53:360–6.

    Article  CAS  PubMed  Google Scholar 

  9. Osada S, Imai H, Tomita H, Tokuyama Y, Okumura N, Matsuhashi N, et al. Serum cytokine levels in response to hepatic cryoablation. J Surg Oncol. 2007;95:491–8.

    Article  CAS  PubMed  Google Scholar 

  10. Poston G. Cryosurgery for colorectal liver metastases. Hepatogastroenterology. 2001;48:323–4.

    CAS  PubMed  Google Scholar 

  11. Hori S, Nomura T, Sakaguchi TJ. Control of regulatory T cell development by the transcription factor Foxp3. Science. 2003;299:1057–61.

    Article  CAS  PubMed  Google Scholar 

  12. Curiel TJ, Coukos G, Zou L, Alvarez X, Cheng P, Mottram P, et al. Specific recruitment of regulatory T cells in ovarian carcinoma fosters immune privilege and predicts reduced survival. Nat Med. 2004;10:942–9.

    Article  CAS  PubMed  Google Scholar 

  13. Sasada T, Kimura M, Yoshida Y, Kanai M, Takabayashi A. CD4+CD25+ regulatory T cells in patients with gastrointestinal malignancies: possible involvement of regulatory T cells in disease progression. Cancer. 2003;98:1089–99.

    Article  PubMed  Google Scholar 

  14. Ormandy LA, Hillemann T, Wedemeyer H, Manns MP, Greten TF, Korangy F. Increased populations of regulatory T cells in peripheral blood of patients with hepatocellular carcinoma. Cancer Res. 2005;65:2457–64.

    Article  CAS  PubMed  Google Scholar 

  15. Stoop JN, van der Molen RG, Baan CC, van der Laan LJ, Kuipers EJ, Kusters JG, et al. Regulatory T cells contribute to the impaired immune response in patients with chronic hepatitis B virus infection. Hepatology. 2005;41:771–8.

    Article  CAS  PubMed  Google Scholar 

  16. Sato E, Olson SH, Ahn J, Bundy B, Nishikawa H, Qian F, et al. Intraepithelial CD8+ tumor infiltrating lymphocytes and a high CD8+/regulatory T cell ratio are associated with favorable prognosis in ovarian cancer. Proc Natl Acad Sci USA. 2005;102:18538–43.

    Article  CAS  PubMed  Google Scholar 

  17. Gao Q, Qiu SJ, Fan J, Zhou J, Wang XY, Xiao YS, et al. Intratumoral balance of regulatory and cytotoxic T cells is associated with prognosis of hepatocellular carcinoma after resection. J Clin Oncol. 2007;25:2586–93.

    Article  PubMed  Google Scholar 

  18. Fu JL, Xu DP, Liu ZW, Shi M, Zhao P, Fu BY, et al. Increased regulatory T cells correlate with CD8+ T cell impairment and poor survival in hepatocellular carcinoma patients. Gastroenterology. 2007;132:2328–39.

    Article  PubMed  Google Scholar 

  19. Kono K, Kawaida H, Takahashi A, Sugai H, Mimura K, Miyagawa N, et al. CD4+CD25high regulatory T cells increase with tumor stage in patients with gastric and esophageal cancers. Cancer Immunol Immunother. 2006;55:1064–71.

    Article  CAS  PubMed  Google Scholar 

  20. Levy MY, Sidana A, Chowdhury WH, Solomon SB, Drake CG, Rodriguez R, et al. Cyclophosphamide unmasks an anti-metastatic effect of local tumor cryoablation. J Pharmacol Exp Ther. 2009;330:596–601.

    Article  CAS  PubMed  Google Scholar 

  21. Llovet JM, Bru C, Bruix J. Prognosis of hepatocellular carcinoma: the BCLC staging classification. Semin Liver Dis. 1999;19:329–38.

    Article  CAS  PubMed  Google Scholar 

  22. Therasse P, Arbuck SG, Eisenhauer EA, Wanders J, Kaplan RS, Rubinstein L, et al. New guidelines to evaluate the response to treatment in solid tumors. European Organization for Research and Treatment of Cancer, National Cancer Institute of the United States, National Cancer Institute of Canada. J Natl Cancer Inst. 2000;92:205–16.

    Article  CAS  PubMed  Google Scholar 

  23. Allan SE, Crome SQ, Crellin NK, Passerini L, Steiner TS, Bacchetta R, et al. Activation-induced FOXP3 in human T effector cells does not suppress proliferation or cytokine production. Int Immunol. 2007;19:345–54.

    Article  CAS  PubMed  Google Scholar 

  24. Baecher-Allan C, Anderson DE. Regulatory cells and human cancer. Semin Cancer Biol. 2006;16:98–105.

    Article  CAS  PubMed  Google Scholar 

  25. Yamaguchi T, Sakaguchi S. Regulatory T cells in immune surveillance and treatment of cancer. Semin Cancer Biol. 2006;16:115–23.

    Article  CAS  PubMed  Google Scholar 

  26. Khazaie K, von Boehmer H. The impact of CD4+CD25+ Treg on tumor specific CD8+ T cell cytotoxicity and cancer. Semin Cancer Biol. 2006;16:124–36.

    Article  CAS  PubMed  Google Scholar 

  27. Sasaki A, Tanaka F, Mimori K, Inoue H, Kai S, Shibata K, et al. Prognostic value of tumor-infiltrating FOXP3+ regulatory T cells in patients with hepatocellular carcinoma. Eur J Surg Oncol. 2008;34:173–9.

    CAS  PubMed  Google Scholar 

  28. Valzasina B, Piconese S, Guiducci C, Colombo MP. Tumor-induced expansion of regulatory T cells by conversion of CD4+ CD25− lymphocytes is thymus and proliferation independent. Cancer Res. 2006;66:4488–95.

    Article  CAS  PubMed  Google Scholar 

  29. Zou WP. Immuno-suppressive networks in the tumor environment and their therapeutic relevance. Nat Rev Cancer. 2005;5:263–74.

    Article  CAS  PubMed  Google Scholar 

  30. Yamazaki S, Iyoda T, Tarbell K, Olson K, Velinzon K, Inaba K, et al. Direct expansion of functional CD25+ CD4+ regulatory T cells by antigen-processing dendritic cells. J Exp Med. 2003;198:235–47.

    Article  CAS  PubMed  Google Scholar 

  31. Van Eden W, Wick G, Albani S, Cohen I. Stress, heat shock proteins, and autoimmunity: how immune responses to heat shock proteins are to be used for the control of chronic inflammatory diseases. Ann N Y Acad Sci. 2007;1113:217–37.

    Article  PubMed  Google Scholar 

  32. Ravindranath MH, Wood TF, Soh D, Gonzales A, Muthugounder S, Perez C, et al. Cryosurgical ablation of liver tumors in colon cancer patients increases the serum total ganglioside level and then selectively augments antiganglioside IgM. Cryobiology. 2002;45:10–21.

    Article  CAS  PubMed  Google Scholar 

  33. Friedline RH, Brown DS, Nguyen H, Kornfeld H, Lee J, Zhang Y, et al. CD4+ regulatory T cells require CTLA-4 for the maintenance of systemic tolerance. J Exp Med. 2009;206:421–34.

    Article  CAS  PubMed  Google Scholar 

  34. Wing K, Onishi Y, Prieto-Martin P, Yamaguchi T, Miyara M, Fehervari Z, et al. CTLA-4 control over Foxp3+ regulatory T cell function. Science. 2008;322:271–5.

    Article  CAS  PubMed  Google Scholar 

  35. Xu D, Fu J, Jin L, Zhang H, Zhou C, Zou Z, et al. Circulating and liver resident CD4+CD25+ regulatory T cells actively influence the antiviral immune response and disease progression in patients with hepatitis B. J Immunol. 2006;177:739–47.

    CAS  PubMed  Google Scholar 

  36. Baecher-Allan C, Brown JA, Freeman GJ, Hafler DA. CD4+CD25 high regulatory cells in human peripheral blood. J Immunol. 2001;167:1245–53.

    CAS  PubMed  Google Scholar 

  37. Poon RT, Fan ST, Ng IO, Lo CM, Liu CL, Wong J. Different risk factors and prognosis for early and late intrahepatic recurrence after resection of hepatocellular carcinoma. Cancer. 2000;89:500–7.

    Article  CAS  PubMed  Google Scholar 

  38. Wang C, Lu Y, Chen Y, Feng Y, An L, Wang X, et al. Prognostic factors and recurrence of hepatitis B-related hepatocellular carcinoma after argon-helium cryoablation: a prospective study. Clin Exp Metastasis. 2009;26:839–48.

    Article  CAS  PubMed  Google Scholar 

  39. Wang ZL, Liang P, Dong BW, Yu XL, de Yu J. Prognostic factors and recurrence of small hepatocellular carcinoma after hepatic resection or microwave ablation: a retrospective study. J Gastrointest Surg. 2008;12:327–37.

    Article  PubMed  Google Scholar 

  40. Poon RT, Lau C, Pang R, Ng KK, Yuen J, Fan ST. High serum vascular endothelial growth factor levels predict poor prognosis after radiofrequency ablation of hepatocellular carcinoma: importance of tumor biomarker in ablative therapies. Ann Surg Oncol. 2007;14:1835–45.

    Article  PubMed  Google Scholar 

  41. Udagawa M, Kudo-Saito C, Hasegawa G, Yano K, Yamamoto A, Yaguchi M, et al. Enhancement of immunologic tumor regression by intratumoral administration of dendritic cells in combination with cryoablative tumor pretreatment and Bacillus Calmette-Guerin cell wall skeleton stimulation. Clin Cancer Res. 2006;12:7465–75.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported in part by the Key Scientific and Technological Research Foundation of the National Special-purpose Program, No. 2008ZX10002-018, and the National Natural Science Foundation of China, No. 30901328.

Author information

Authors and Affiliations

  1. Center of Therapeutic Research for Hepatocellular Carcinoma, Beijing 302 Hospital, Beijing, 100039, China

    Lin Zhou, Yin-Ying Lu, Chun-Ping Wang, Lin-Jing An, Xin-Zhen Wang, Zhen Zeng & Yong-Ping Yang

  2. Research Center for Biological Therapy, Beijing 302 Hospital, Beijing, 100039, China

    Jun-Liang Fu, Bao-Yun Fu, Chun-Bao Zhou & Fu-Sheng Wang

Authors
  1. Lin Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jun-Liang Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yin-Ying Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Bao-Yun Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chun-Ping Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Lin-Jing An
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xin-Zhen Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Zhen Zeng
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Chun-Bao Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Yong-Ping Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Fu-Sheng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yong-Ping Yang.

Additional information

L. Zhou and J.-L. Fu contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhou, L., Fu, JL., Lu, YY. et al. Regulatory T cells are associated with post-cryoablation prognosis in patients with hepatitis B virus-related hepatocellular carcinoma. J Gastroenterol 45, 968–978 (2010). https://doi.org/10.1007/s00535-010-0243-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00535-010-0243-3

Keywords

Search

Navigation